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  1. Article ; Online: Color-Coded Super-Resolution Small-Molecule Imaging.

    Beuzer, Paolo / La Clair, James J / Cang, Hu

    Chembiochem : a European journal of chemical biology

    2016  Volume 17, Issue 11, Page(s) 999–1003

    Abstract: Although the development of super-resolution microscopy dates back to 1994, its applications have been primarily focused on visualizing cellular structures and targets, including proteins, DNA and sugars. We now report on a system that allows both ... ...

    Abstract Although the development of super-resolution microscopy dates back to 1994, its applications have been primarily focused on visualizing cellular structures and targets, including proteins, DNA and sugars. We now report on a system that allows both monitoring of the localization of exogenous small molecules in live cells at low resolution and subsequent super-resolution imaging by using stochastic optical reconstruction microscopy (STORM) on fixed cells. This represents a powerful new tool to understand the dynamics of subcellular trafficking associated with the mode and mechanism of action of exogenous small molecules.
    MeSH term(s) Antibodies, Monoclonal/chemistry ; Antibodies, Monoclonal/metabolism ; Carbocyanines/chemistry ; Cell Line, Tumor ; Color ; DNA/chemistry ; DNA/metabolism ; Humans ; Microscopy, Fluorescence ; Proteins/chemistry ; Proteins/metabolism ; Small Molecule Libraries/chemistry
    Chemical Substances Alexa Fluor 647 ; Antibodies, Monoclonal ; Carbocyanines ; Proteins ; Small Molecule Libraries ; DNA (9007-49-2)
    Language English
    Publishing date 2016-04-26
    Publishing country Germany
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2020469-3
    ISSN 1439-7633 ; 1439-4227
    ISSN (online) 1439-7633
    ISSN 1439-4227
    DOI 10.1002/cbic.201600013
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Establishment of a replication fork barrier following induction of DNA binding in mammalian cells.

    Beuzer, Paolo / Quivy, Jean-Pierre / Almouzni, Geneviève

    Cell cycle (Georgetown, Tex.)

    2014  Volume 13, Issue 10, Page(s) 1607–1616

    Abstract: Understanding the mechanisms that lead to replication fork blocks (RFB) and the means to bypass them is important given the threat that they represent for genome stability if inappropriately handled. Here, to study this issue in mammals, we use ... ...

    Abstract Understanding the mechanisms that lead to replication fork blocks (RFB) and the means to bypass them is important given the threat that they represent for genome stability if inappropriately handled. Here, to study this issue in mammals, we use integrated arrays of the LacO and/or TetO as a tractable system to follow in time a process in an individual cell and at a single locus. Importantly, we show that induction of the binding by LacI and TetR proteins, and not the presence of the repeats, is key to form the RFB. We find that the binding of the proteins to the arrays during replication causes a prolonged persistence of replication foci at the site. This, in turn, induces a local DNA damage repair (DDR) response, with the recruitment of proteins involved in double-strand break (DSB) repair such as TOPBP1 and 53BP1, and the phosphorylation of H2AX. Furthermore, the appearance of micronuclei and DNA bridges after mitosis is consistent with an incomplete replication. We discuss how the many DNA binding proteins encountered during replication can be dealt with and the consequences of incomplete replication. Future studies exploiting this type of system should help analyze how an RFB, along with bypass mechanisms, are controlled in order to maintain genome integrity.
    MeSH term(s) Animals ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Cell Line, Tumor ; DNA Breaks, Double-Stranded ; DNA Repair ; DNA Replication ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Histones/genetics ; Histones/metabolism ; Humans ; Intracellular Signaling Peptides and Proteins/genetics ; Intracellular Signaling Peptides and Proteins/metabolism ; Mice ; Nuclear Proteins/genetics ; Nuclear Proteins/metabolism ; Phosphorylation ; S Phase ; Tumor Suppressor p53-Binding Protein 1
    Chemical Substances Carrier Proteins ; DNA-Binding Proteins ; H2AX protein, human ; Histones ; Intracellular Signaling Peptides and Proteins ; Nuclear Proteins ; TOPBP1 protein, human ; TP53BP1 protein, human ; Tumor Suppressor p53-Binding Protein 1
    Language English
    Publishing date 2014-03-25
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2146183-1
    ISSN 1551-4005 ; 1538-4101 ; 1554-8627
    ISSN (online) 1551-4005
    ISSN 1538-4101 ; 1554-8627
    DOI 10.4161/cc.28627
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 5881: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (2.OG)
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Single dish gradient screening of small molecule localization.

    Beuzer, Paolo / Axelrod, Joshua / Trzoss, Lynnie / Fenical, Willam / Dasari, Ramesh / Evidente, Antonio / Kornienko, Alexander / Cang, Hu / La Clair, James J

    Organic & biomolecular chemistry

    2016  Volume 14, Issue 35, Page(s) 8241–8245

    Abstract: Understanding trafficking in cells and tissues is one of the most critical steps in exploring the mechanisms and modes of action (MOAs) of a small molecule. Typically, deciphering the role of concentration presents one of the most difficult challenges ... ...

    Abstract Understanding trafficking in cells and tissues is one of the most critical steps in exploring the mechanisms and modes of action (MOAs) of a small molecule. Typically, deciphering the role of concentration presents one of the most difficult challenges associated with this task. Herein, we present a practical solution to this problem by developing concentration gradients within single dishes of cells. We demonstrate the method by evaluating fluorescently-labelled probes developed from two classes of natural products that have been identified as potential anti-cancer leads by STORM super-resolution microscopy.
    Language English
    Publishing date 2016-08-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 2097583-1
    ISSN 1477-0539 ; 1477-0520
    ISSN (online) 1477-0539
    ISSN 1477-0520
    DOI 10.1039/c6ob01418f
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Molecular organization of mammalian meiotic chromosome axis revealed by expansion STORM microscopy.

    Xu, Huizhong / Tong, Zhisong / Ye, Qing / Sun, Tengqian / Hong, Zhenmin / Zhang, Lunfeng / Bortnick, Alexandra / Cho, Sunglim / Beuzer, Paolo / Axelrod, Joshua / Hu, Qiongzheng / Wang, Melissa / Evans, Sylvia M / Murre, Cornelis / Lu, Li-Fan / Sun, Sha / Corbett, Kevin D / Cang, Hu

    Proceedings of the National Academy of Sciences of the United States of America

    2019  Volume 116, Issue 37, Page(s) 18423–18428

    Abstract: During prophase I of meiosis, chromosomes become organized as loop arrays around the proteinaceous chromosome axis. As homologous chromosomes physically pair and recombine, the chromosome axis is integrated into the tripartite synaptonemal complex (SC) ... ...

    Abstract During prophase I of meiosis, chromosomes become organized as loop arrays around the proteinaceous chromosome axis. As homologous chromosomes physically pair and recombine, the chromosome axis is integrated into the tripartite synaptonemal complex (SC) as this structure's lateral elements (LEs). While the components of the mammalian chromosome axis/LE-including meiosis-specific cohesin complexes, the axial element proteins SYCP3 and SYCP2, and the HORMA domain proteins HORMAD1 and HORMAD2-are known, the molecular organization of these components within the axis is poorly understood. Here, using expansion microscopy coupled with 2-color stochastic optical reconstruction microscopy (STORM) imaging (ExSTORM), we address these issues in mouse spermatocytes at a resolution of 10 to 20 nm. Our data show that SYCP3 and the SYCP2 C terminus, which are known to form filaments in vitro, form a compact core around which cohesin complexes, HORMADs, and the N terminus of SYCP2 are arrayed. Overall, our study provides a detailed structural view of the meiotic chromosome axis, a key organizational and regulatory component of meiotic chromosomes.
    MeSH term(s) Animals ; Cell Cycle Proteins/metabolism ; Chromosomes, Mammalian/chemistry ; Chromosomes, Mammalian/metabolism ; DNA-Binding Proteins/metabolism ; Male ; Mammals/genetics ; Meiosis ; Mice ; Microscopy/methods ; Spermatocytes/metabolism ; Staining and Labeling ; Synaptonemal Complex/metabolism
    Chemical Substances Cell Cycle Proteins ; DNA-Binding Proteins ; HORMAD2 protein, mouse ; Nohma protein, mouse ; Sycp2 protein, mouse ; Sycp3 protein, mouse
    Language English
    Publishing date 2019-08-23
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.1902440116
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  5. Article ; Online: Ritterostatin G

    Ambrose, Andrew J / Santos, Evelyne A / Jimenez, Paula C / Rocha, Danilo D / Wilke, Diego V / Beuzer, Paolo / Axelrod, Josh / Kumar Kanduluru, Ananda / Fuchs, Philip L / Cang, Hu / Costa-Lotufo, Letícia V / Chapman, Eli / La Clair, James J

    Chembiochem : a European journal of chemical biology

    2017  Volume 18, Issue 6, Page(s) 506–510

    Abstract: Natural products discovered by using agnostic approaches, unlike rationally designed leads or those obtained through high-throughput screening, offer the ability to reveal new biological pathways and, hence, serve as an important vehicle to unveil new ... ...

    Abstract Natural products discovered by using agnostic approaches, unlike rationally designed leads or those obtained through high-throughput screening, offer the ability to reveal new biological pathways and, hence, serve as an important vehicle to unveil new avenues in drug discovery. The ritterazine-cephalostatin family of natural products displays robust and potent antitumor activities, with sub-nanomolar growth inhibition against multiple cell lines and potent activity in xenograft models. Herein, we used comparative cellular and molecular biological methods to uncover the ritterazine-cephalostatin cytotoxic mode of action (MOA) in human tumor cells. Our findings indicated that, whereas ritterostatin G
    MeSH term(s) Cell Line, Tumor ; Cell Proliferation/drug effects ; Cell Survival/drug effects ; Cells, Cultured ; Coumarins/chemistry ; Drug Delivery Systems ; Endoplasmic Reticulum Chaperone BiP ; Heat-Shock Proteins/metabolism ; Humans ; Molecular Probes ; Molecular Structure ; Phenazines/chemistry ; Protein Binding/drug effects ; Pyrazines/chemistry ; Pyrazines/pharmacology ; Steroids/chemistry
    Chemical Substances Coumarins ; Endoplasmic Reticulum Chaperone BiP ; HSPA5 protein, human ; Heat-Shock Proteins ; Molecular Probes ; Phenazines ; Pyrazines ; Steroids ; ritterostatin GN1N
    Language English
    Publishing date 2017-02-02
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2020469-3
    ISSN 1439-7633 ; 1439-4227
    ISSN (online) 1439-7633
    ISSN 1439-4227
    DOI 10.1002/cbic.201600669
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Use of optical tweezers technology for long-term, focal stimulation of specific subcellular neuronal compartments.

    D'Este, Elisa / Baj, Gabriele / Beuzer, Paolo / Ferrari, Enrico / Pinato, Giulietta / Tongiorgi, Enrico / Cojoc, Dan

    Integrative biology : quantitative biosciences from nano to macro

    2011  Volume 3, Issue 5, Page(s) 568–577

    Abstract: Spatial regulation of secretory molecule release is a sophisticated mechanism used by the nervous system to control network development and finely tune the activity of each synapse. Great efforts have been made to develop techniques that mimic secretory ... ...

    Abstract Spatial regulation of secretory molecule release is a sophisticated mechanism used by the nervous system to control network development and finely tune the activity of each synapse. Great efforts have been made to develop techniques that mimic secretory molecule release with the aim of stimulating neurons as close as possible to physiological conditions. However, current techniques have poor spatial resolution or low flexibility. Here, we propose a novel approach to achieve focal and prolonged stimulation of neurons using optical tweezers and single microbeads functionalized with a secretory molecule, the neurotrophin brain-derived neurotrophic factor (BDNF). BDNF is a key regulator of neuronal development and plasticity. We show that single BDNF-coated microbeads can be extracted with optical tweezers from small reservoirs and positioned with submicrometric precision to specific sites on the dendrites of cultured hippocampal neurons. Localized contact of microbeads functionalized with BDNF, but not with bovine serum albumin (BSA), induced focal increase of calcium signaling in the stimulated dendrite, specific activation of the TrkB receptor pathway and influenced the development of growth cones. Remarkably, a single BDNF-coated bead localized on a dendrite was found to be enough for TrkB phosphorylation, an efficient and long-lasting activation of calcium signaling in the soma, and c-Fos signaling in the nucleus, comparable to bath stimulation conditions. These findings support the use of optical tweezer technology for long-term, localized stimulation of specific subcellular neuronal compartments.
    MeSH term(s) Animals ; Brain-Derived Neurotrophic Factor/administration & dosage ; Cell Line ; Neurons/drug effects ; Neurons/physiology ; Optical Tweezers ; Physical Stimulation/methods ; Rats ; Subcellular Fractions/drug effects ; Subcellular Fractions/physiology
    Chemical Substances Brain-Derived Neurotrophic Factor
    Language English
    Publishing date 2011-05
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2480063-6
    ISSN 1757-9708 ; 1757-9694
    ISSN (online) 1757-9708
    ISSN 1757-9694
    DOI 10.1039/c0ib00102c
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: Use of optical tweezers technology for long-term, focal stimulation of specific subcellular neuronal compartments

    D'Este, Elisa / Baj, Gabriele / Beuzer, Paolo / Ferrari, Enrico / Pinato, Giulietta / Tongiorgi, Enrico / Cojoc, Dan

    Integrative biology. 2011 May 3, v. 3, no. 5

    2011  

    Abstract: Spatial regulation of secretory molecule release is a sophisticated mechanism used by the nervous system to control network development and finely tune the activity of each synapse. Great efforts have been made to develop techniques that mimic secretory ... ...

    Abstract Spatial regulation of secretory molecule release is a sophisticated mechanism used by the nervous system to control network development and finely tune the activity of each synapse. Great efforts have been made to develop techniques that mimic secretory molecule release with the aim of stimulating neurons as close as possible to physiological conditions. However, current techniques have poor spatial resolution or low flexibility. Here, we propose a novel approach to achieve focal and prolonged stimulation of neurons using optical tweezers and single microbeads functionalized with a secretory molecule, the neurotrophin brain-derived neurotrophic factor (BDNF). BDNF is a key regulator of neuronal development and plasticity. We show that single BDNF-coated microbeads can be extracted with optical tweezers from small reservoirs and positioned with submicrometric precision to specific sites on the dendrites of cultured hippocampal neurons. Localized contact of microbeads functionalized with BDNF, but not with bovine serum albumin (BSA), induced focal increase of calcium signaling in the stimulated dendrite, specific activation of the TrkB receptor pathway and influenced the development of growth cones. Remarkably, a single BDNF-coated bead localized on a dendrite was found to be enough for TrkB phosphorylation, an efficient and long-lasting activation of calcium signaling in the soma, and c-Fos signaling in the nucleus, comparable to bath stimulation conditions. These findings support the use of optical tweezer technology for long-term, localized stimulation of specific subcellular neuronal compartments.
    Keywords bovine serum albumin ; calcium signaling ; dendrites ; microbeads ; optical traps ; phosphorylation ; synapse
    Language English
    Dates of publication 2011-0503
    Size p. 568-577.
    Publishing place The Royal Society of Chemistry
    Document type Article
    ZDB-ID 2480063-6
    ISSN 1757-9708 ; 1757-9694
    ISSN (online) 1757-9708
    ISSN 1757-9694
    DOI 10.1039/c0ib00102c
    Database NAL-Catalogue (AGRICOLA)

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